反应详情

Isomeric preference of the mRPE65/tLRAT mediated esterification of retinols: The effect of mRPE65 on the processing of 11-cis-and all-trans-retinols was determined by monitoring the formation of tLRAT catalyzed retinyl esters from added all-trans-retinol [11,12-3H2], 11-cis-retinol [15-3H] and mRPE65. The reaction mixture (volume 0.1 mL) contains 100 mM Tris (pH 8.4), 5 μM of tLRAT 0.3% CHAPS, 1 mM dithiothreitol and 0.2 μm of all-trans-retinol [11,12-3H2] or 11-cis-retinol [15-3H] and mRPE65 (0.02 μM) or 200 μM/0.4% DPPC/BSA was incubated for 10 min at room temperature. After 10 min the reaction was quenched with 500 μL methanol, 100 μL of water and 500 μL of hexane. The amount of retinyl palmitate formed, as determined by normal phase-HPLC, was used as a measure of activity. Each experiment was done in triplicate, and the data points used are an average of these three points. The standard error was presented as error bars.

1. customAfter 10 min the reaction was quenched with 500 μL methanol, 100 μL of water and 500 μL of hexane